Mechanisms of Anticancer Activity of Sulforaphane from Brassica oleracea in HEp-2 Human Epithelial Carcinoma Cell Line
Mechanisms of Anticancer Activity of Sulforaphane from Brassica oleracea in HEp-2 Human Epithelial Carcinoma Cell Line
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DOI：
10.7314/apjcp.2012.13.5.2095
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发表时间：
2012-01-01
影响因子：
--
通讯作者：
Thangam, E. Berla
中科院分区：
文献类型：
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作者：
Devi, J. Renuka;Thangam, E. Berla
萝卜硫素（SFN）是一种存在于甘蓝科植物中的异硫氰酸酯，具有抗癌和抗氧化活性。在这项研究中，我们从红甘蓝（甘蓝变种红）分离SFN和评估比较不同的馏分（标准SFN，提取和纯化SFN）的抗增殖活性通过MTT法在人上皮癌HEp-2和Vero细胞。还研究了通过p53、Bax和bcl-2介导的可能的细胞凋亡机制。通过HPLC收集SFN级分，富集其SFN含量并确认。Western blotting和RT PCR检测细胞凋亡相关蛋白的表达。结果显示，发现标准SFN和纯化SFN浓度具有更接近的IC 50，其等于58.96 μ g/ml（HEp-2细胞）、61.2 μ g/ml（Vero细胞），并且小于发现的提取物113 μ g/ml（HEp-2细胞）和125 μ g/ml（Vero细胞）。进一步的凋亡机制研究表明，纯化的SFN下调bcl-2（抗凋亡）的表达，而上调p53和Bax（促凋亡）蛋白，以及caspase-3。本研究表明，纯化的SFN具有与Std SFN相同的抗增殖作用，其诱导HEp-2细胞凋亡的机制可能与p53诱导、bax和bcl-2信号通路有关。
Sulforaphane (SFN) an isothiocyanate formed by hydrolysis of glucosinolates found in Brassica oleraceae is reported to possess anticancer and antioxidant activities. In this study, we isolated SFN from red cabbage (Brassica oleraceae var rubra) and evaluated the comparative antiproliferative activity of various fractions (standard SFN, extract and purified SFN) by MTT assay in human epithelial carcinoma HEp -2 and and Vero cells. Probable apoptotic mechanisms mediated through p53, bax and bcl-2 were also examined. The SFN fraction was collected by HPLC, enriched for its SFN content and confirmed. Expression of apoptosis-related proteins was detected by western blotting and RT PCR. Results showed that Std SFN and purified SFN concentration found to have closer IC50 which is equal to 58.96 microgram/ml (HEp-2 cells), 61.2 microgram/ml (Vero cells) and less than the extract which is found to be 113 microgram/ml (HEp-2 cells) and 125 microgram/ml (Vero cells). Further studies on apoptotic mechanisms showed that purified SFN down-regulated the expression of bcl-2 (antiapoptotic), while up-regulating p53 and Bax (proapoptotic) proteins, as well as caspase-3. This study indicates that purified SFN possesses antiproliferative effects the same as Std SFN and its apoptotic mechanism in HEp-2 cells could be mediated through p53 induction, bax and bcl-2 signaling pathways.